Volume 252, 2024

Designing Michaelases: exploration of novel protein scaffolds for iminium biocatalysis

Abstract

Biocatalysis is becoming a powerful and sustainable alternative for asymmetric catalysis. However, enzymes are often restricted to metabolic and less complex reactivities. This can be addressed by protein engineering, such as incorporating new-to-nature functional groups into proteins through the so-called expansion of the genetic code to produce artificial enzymes. Selecting a suitable protein scaffold is a challenging task that plays a key role in designing artificial enzymes. In this work, we explored different protein scaffolds for an abiological model of iminium-ion catalysis, Michael addition of nitromethane into E-cinnamaldehyde. We studied scaffolds looking for open hydrophobic pockets and enzymes with described binding sites for the targeted substrate. The proteins were expressed and variants harboring functional amine groups – lysine, p-aminophenylalanine, or N6-(D-prolyl)-L-lysine – were analyzed for the model reaction. Among the newly identified scaffolds, a thermophilic ene-reductase from Thermoanaerobacter pseudethanolicus was shown to be the most promising biomolecular scaffold for this reaction.

Graphical abstract: Designing Michaelases: exploration of novel protein scaffolds for iminium biocatalysis

  • This article is part of the themed collection: Biocatalysis

Associated articles

Supplementary files

Article information

Article type
Paper
Submitted
11 mar. 2024
Accepted
18 mar. 2024
First published
18 mar. 2024
This article is Open Access
Creative Commons BY license

Faraday Discuss., 2024,252, 279-294

Designing Michaelases: exploration of novel protein scaffolds for iminium biocatalysis

A. Gran-Scheuch, S. Hanreich, I. Keizer, J. W. Harteveld, E. Ruijter and I. Drienovská, Faraday Discuss., 2024, 252, 279 DOI: 10.1039/D4FD00057A

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